METHOD FOR DEVELOPING IMPROVED GENE INHIBITORS

这项建议的目标是开发一种通用的方法，快速获得任何感兴趣的基因的有效抑制剂。实现这一目标需要两个步骤：1)在mRNA上选择一个可接近的位置，2)设计一个有效的分子，它将识别并结合到该位置，或通过一些其他方法使其失活。目前，现有的反义技术和可获得的靶点选择仍处于起步阶段，不可靠。因此，我们提出了一种新颖而通用的方法，用于快速开发一种有效的技术来定位任何RNA中的可访问靶点，并针对这些已定位的可访问位点设计有效的抑制剂。我们通过使用反义序列的随机文库来实现这一点，这些反义序列在目标处于自然构象的细胞内发挥作用，自动识别任何所需的目标RNA中最容易接近的位置。通过使用这些识别的可访问位点，我们将设计我们专有的、基于拓扑的反义设计-RNA挂锁，以定向抑制任何所需基因的mRNA。拟议的商业应用：反义技术目前受到以下因素的阻碍：寡核苷酸与靶mRNAs上的大多数位点结合的能力很差，而且很难识别它们可以结合的那些位点。通过克服这一障碍，我们将帮助实现反义技术最初承诺的合理药物设计和药物靶标验证的巨大可能性。

The goal of this proposal is to develop a universal method for rapidly deriving effective inhibitors for any gene of interest. Accomplishing this goal requires two steps: 1) selecting an accessible site on the mRNA, and 2) designing an effective molecule that will recognize that site and bind to it, or inactivate it by some other means. Currently available antisense technology and accessible target site selection is still in its infancy and unreliable. Therefore we propose a novel and general approach for rapidly developing an effective technology for mapping accessible target sites in any RNA as well as engineering effective inhibitors to those mapped accessible sites. We accomplish this by using random libraries of antisense sequences that, functioning inside the cell where the target is in its natural conformation, automatically identify the most accessible sites in any desired target RNA. By employing those identified accessible sites, we will engineer our proprietary, topology-based antisense design, RNA Padlocks, for targeted inhibition of mRNA of any desired gene. PROPOSED COMMERCIAL APPLICATIONS: Antisense technologies are currently hindered by the poor ability of oligonucleotides to bind to most sites on target mRNAs, and the difficulty of identifying those sites to which they can bind. By overcoming this hurdle, we will help realize the great possibilities of rational drug design and drug target validation that antisense technology originally promised.